Mechanism-based inactivation of L-aspartase from Escherichia coli.
Mechanism-based inactivation of L-aspartase from Escherichia coli.
复制标题
基于机制的大肠杆菌 L-天冬氨酸酶失活。
DOI：
10.1021/bi00197a042
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发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Viola,RE
中科院分区：
文献类型：
--
作者：
Schindler,JF;Viola,RE
材料和方法材料。L-天冬氨酸、富马酸、α-甲基-DL-天冬氨酸和L-烯丙基甘氨酸均从Sigma中分离得到，无需进一步纯化即可使用。L-甘氨酸是由L-烯丙基甘氨酸经臭氧氧化制备而成，并按公开发表的方法(Black&Wright，1955)进行纯化。FAA是由20g巴豆酸甲酯在100mL二恶烷中回流4h与13g SEC>；2回流制备的(Bohlman&Inhoffen，1956)。用这种试剂处理会导致烯丙基碳选择性氧化成醛(Trachtenberg，1969)。甘油中无氨的谷氨酸脱氢酶(GDH)来自勃林格，2，4-二内苯肼(DNPH)来自Aldrich。L-天冬氨酸酶是用Karsten等人(1985)的方法提纯的。
MATERIALS AND METHODSMaterials. L-Asparticacid, fumarate, a-methyl-DL-aspartate (AMA), and L-allylglycine were all obtained from Sigma and were used without further purification. l-ASA was prepared by the ozonolysis of l-allylglycine and was purified according to a published method (Black & Wright, 1955). FAA was prepared by refluxing 20 g of methyl crotonate in 100 mL of dioxane for 4 h with 13 g of SeC> 2 (Bohlman & Inhoffen, 1956). Treatment with this reagent leads to the selective oxidation of the allylic carbon to the aldehyde (Trachtenberg, 1969). Glutamate dehydrogenase (GDH), ammonia-free in glycerol was from Boehringer, and 2, 4-dintrophenylhydrazine (DNPH) was from Aldrich. The enzyme L-aspartase was purified by theprocedure of Karsten et al.(1985).